I am currently pursuing a study that involves the exposure of a breast cancer cell line to a particular cytotoxic substance that may/may not alter its morphology. Is there currently any protocol for the examination of breast cancer cell morphology? Or is there a reliable source that I could compare observations with? Thank you. Do you have any help would be greatly appreciated.
Dear Jason
Elaborate what exactly are u looking at with respect to cell morphology.
according to my knowledge, nuclear stains like Propidium Iodide and DAPI can be used to check if the cell is shrinken or the dispersion of nuclear material in the medium as the stains are DNA binding dyes. (Instrument required is High content screening system) -- For live dead assay these two dyes are prominent.
If not there are many fluorescent cell surface/nuclear membrane/mitochondrial membrane specific commercial markers which u can use for treated and untreated cells and check for morphology.
So based on your specification with respect to cell morphology you can look for commercially available antibody conjugated dyes specific to specific membrane or cell component and stain them followed by viewing them in High content screening system.
I hope i can be more specific after you under stand my query.
You can use Phallodin staining which stains the actin filaments to observe the morphology change of the cell after drug treatment.
I have some suggestion for you. So many different breast cancer cell lines are available from ATCC. For example MCF-7, MDAMB-468, etc. The company describes the morphology and growth characteristics of each of them which may vary from each other. The best protocol is to read the company protocol carefully and check through inverted microscope the morphology and examine it, i.c. whether the morphology matched with the said protocol. After that you may stain with any of the above stain as mentioned above. When you are adding a drug, check the morphology again through inverted microscope. If the cells are round shaped the drug may have severe effect and cells may be pre-apoptotic or necrotic. If the cells are completely detached from the plate or flask the cells may be already dead. So, inverted microscope with attached camera may give you a lots of information. If you think there is a toxicity effect of the drugs you can check by MTT assay.
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